Imagen de Portada

Characterization of a sesquiterpene synthase and a short-chain dehydrogenase in zerumbone biosynthesis and the applications in engineered Saccharomyces cerevisiae

Guardado en:
Publicado en:Frontiers in Plant Science vol. 16 (Oct 2025), p. 1635141-1635153
Autor principal: Xu, Mengdie
Otros Autores: Xia, Yimeng, Fang, Gaowei, Li, Tangli, Ma, Jing, Li, Dengyu, Qiuhui Wei, Tu, Lichan, Yin, Xiaopu, Engineering Laboratory of Development and Application of Traditional Chinese Medicines, Collaborative Innovation Center of Traditional Chinese Medicines of Zhejiang Province, Hangzhou Normal University, Hangzhou, China"], Hu, Tianyuan
Publicado:
Frontiers Media SA
Materias:
Transcriptomes
Biosynthesis
Software
Caryophyllene
Metabolic engineering
E coli
Metabolism
Yeast
Genes
Dehydrogenases
Factories
Physicochemical properties
Sesquiterpenes
Dehydrogenase
Oxidation
Molecular docking
Site-directed mutagenesis
Cloning
Zerumbone
DNA polymerase
Precursors
Humulene
Sesquiterpene synthase
Enzymes
Saccharomyces cerevisiae
Environmental
Acceso en línea:Citation/Abstract
Full Text
Full Text - PDF
Etiquetas: Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
Resumen:IntroductionZerumbone is a pharmacologically active sesquiterpenoid with limited availability. This study aims to elucidate its biosynthetic pathway in Curcuma wenyujin by identifying and characterizing the key enzymes responsible for its production.MethodsCandidate genes were selected via transcriptome analysis and phylogenetics. CwTPS8 and CwSDR1 were cloned and functionally characterized using in vitro enzyme assays and heterologous expression in engineered Saccharomyces cerevisiae . Molecular docking and site-directed mutagenesis were applied to investigate the catalytic mechanism of CwTPS8.ResultsCwTPS8 was identified as a multifunctional sesquiterpene synthase that catalyzes the formation of α-humulene (a key zerumbone precursor) and β-caryophyllene as main products, along with several minor sesquiterpenes. Mutagenesis studies identified critical residues (e.g., Thr437, Cys436) that significantly shift product specificity toward α-humulene. CwSDR1 was characterized as a short-chain dehydrogenase that efficiently oxidizes 8-hydroxy-α-humulene to zerumbone. A de novo biosynthetic pathway was reconstructed in yeast, resulting in zerumbone production at 0.50 μg/L.DiscussionThis study expands the genetic toolkit for zerumbone biosynthesis and provides insights into enzyme engineering and metabolic engineering strategies to enhance production. Limitations in precursor supply and catalytic efficiency highlight areas for future optimization.
ISSN:1664-462X
DOI:10.3389/fpls.2025.1635141
Fuente:Agriculture Science Database